Glioblastoma (GB) is the most common primary brain tumor in adults and has a dismal prognosis, which is attributed largely to extreme heterogeneity in the cells that make up the cancer, continual evolution of molecular and genetic alterations, and thus resistance of the tumor to conventional treatments. Beyond surgical resection, systemic treatment with cytotoxic compounds is the main treatment for patients with solid tissue tumors. Traditionally, human glioma tumors have been grown ex vivo in the presence of high concentrations of serum as adherent cultures. While these serum grown cultures can be maintained for extensive periods of time and the resulting progeny are tumorigenic in animal models, they do not display many of the defining phenotypic features that are seen in the primary tumor. Recently, application of the Neurosphere Assay [NSA], which has been used to isolate, expand and study neural stem cells, has been applied to GB. Under these conditions, GB cell lines can be generated under defined conditions and importantly following implantation of the cultured cells; the resulting tumors exhibit the classic in vivo features of human GB; recapitulating the morphology, genotype and gene expression patterns of primary GBMs, as well as having extensive migratory and infiltrative capacity. Hence, application of the NSA to the study of GB may provide a more biologically relevant model system for studying glioma biology and maybe exploited for identification of new therapeutic agents. Currently there are no commercial sources of NSA isolated GB cell lines that have been extensively characterized both phenotypically [in vitro and in vivo] and genomically. Within this application we are proposing to isolate and bank 50 such patient derived lines and make them available to the research community.